Dynamic light scattering based microrheology of complex fluids with improved single-scattering mode detection

ABSTRACT

A fluid characterization measuring instrument is disclosed that comprises a sample vessel for a bulk complex sample fluid having a capacity that is substantially larger than a domain size of the complex sample fluid and that is sufficiently large to cause bulk scattering effects to substantially exceed surface effects for the complex fluid sample, a coherent light source positioned to illuminate the bulk complex sample fluid in the sample vessel and a first fibre having a first end positioned to receive backscattered light from the sample after it has interacted with the sample. The first fibre can also be positioned close enough to an optical axis of the coherent light source and to the sample vessel to substantially decrease a contribution of multiply scattered light in the backscattered light. The instrument can further comprise a first photon-counting detector positioned to receive the backscattered light from a second end of the fibre, correlation logic responsive to the first photon-counting detector and single-scattering fluid property analysis logic responsive to the correlation logic and operative to derive at least one fluid property for the sample fluid.

This application is a continuation of application Ser. No. 15/013,084,filed Feb. 2, 2016, which is a continuation of application Ser. No.13/390,915, filed Sep. 13, 2012, which is a US National Phasecounterpart of international application number PCT/GB2010/51354, filedAug. 17, 2010, which claims priority to U.S. provisional application No.61/274,480, filed Aug. 17, 2009, which are all herein incorporated byreference.

FIELD OF THE INVENTION

This invention relates to methods and apparatus for obtaining theviscoelastic parameters of complex fluids, such as colloidal andbiological complex fluids.

BACKGROUND OF THE INVENTION

Viscoelasticity means the simultaneous existence of viscous and elasticproperties in a material. Many complex and structured fluids exhibitviscoelastic characteristics, i.e., they have the ability to both storeenergy like an elastic solid as well as dissipate energy such as aviscous liquid. When a stress is applied to such a viscoelastic fluid itstores some of the energy input, instead of dissipating all of it asheat and it may recover part of its deformation when the stress isremoved.

The elastic modulus or G′ represents storage of elastic energy, whilethe loss modulus G″ represents the viscous dissipation of that energy.The magnitude of G′ and G″ for most complex fluids depends upon the timescales or frequency at which the property is probed. Depending upon thestress relaxation mechanisms present in the complex fluids, they mayexhibit different behaviour (either G′>G″ or G″>G′ or G′=G″) atdifferent frequencies. Having the ability to probe the viscoelasticresponse over a wide frequency range therefore provides insights intothe stress-relaxation mechanisms in complex fluids, and since this isconnected to the underlying structure of the complex fluid, insightsinto the underlying structure can be obtained.

Currently, high end rotational rheometers are used to measure theseviscoelastic properties, but the measurement time can be quite longdepending upon the frequency being probed. Also, a considerable amountof time can be spent in cleaning the rheometer's stage and preparing thetest before the next sample can be loaded, making high-throughputmeasurements quite challenging. Other disadvantages of rotationalrheometers include that they provide access to a very limited frequencyrange, and they require large sample volumes, typically greater than 1ml.

Optical-based Microrheological techniques have also been used to measureviscoelastic properties of complex fluids. These involve embedding probeparticles into a viscoelastic fluid of interest (polymer solution,surfactant solution etc.) and following the thermal motion of the probeparticles. The thermally driven random motion of colloidal spheressuspended in a complex fluid is very different than the diffusiveBrownian motion of similar spheres suspended in a purely viscous fluid(e.g simple Newtonian fluid). When suspended in complex fluids, whichexhibit elasticity, the probe particles exhibit sub diffusive motion orif the elasticity becomes very significant the probe particles maybecome locally bound. As the microstructure slowly relaxes, it allowsthe particles to escape this elastic ‘cage.’ This motion of probeparticles as a function of time can be obtained from mean squareddisplacement <Δr²(t)> of probe particles which can be obtained from theelectric field autocorrelation function obtained from a Dynamic LightScattering (DLS) experiment:

g ⁽¹⁾(τ)=exp(−⅙q ² Δr ²(r))

Once the mean squared displacement, <Δr²(t)> is obtained, it can berelated through to the complex viscoelastic modulus G* and through tothe elastic G′ and viscous modulus G″ through:

${{G^{\prime}(\omega)} = {{{G^{*}(\omega)}}{\cos \left( {{{\pi\alpha}(\omega)}/2} \right)}}},{{G^{''}(\omega)} = {{{G^{*}(\omega)}}{\sin \left( {\pi \; {{\alpha (\omega)}/2}} \right)}}},{{{where}{{G^{*}(\omega)}}} \approx {\frac{k_{B}T}{\pi \; a{\langle{\Delta \; {r^{2}\left( {1/\omega} \right)}}\rangle}{\Gamma \left\lbrack {1 + {\alpha (\omega)}} \right\rbrack}}.}}$

This analysis is based on two key assumptions:

-   -   The system exhibits single scattering. As the system becomes        multiply scattering the analysis no longer remains valid.    -   The scattering is dominated by the embedded probe particles, as        the whole principle is based on following the motion of the        embedded probe particles.

Many complex fluids at even moderate concentrations start to contributequite significantly to the scattered light signal. In order to ensurethe domination of the scattering by probe particles, they need to beadded in moderately high concentrations (but still much less than 0.5vol %). Adding probe particles in these moderately concentrated regimesmakes the system quite turbid and multiple scattering tends to becomevery significant.

In these types of systems, the concentration of probe particles can beraised even further to enter into the strongly multiply scatteringregime, while changing the analysis from that described above totheories developed for using the multiply scattered light in themicrorheological analysis. This then evolves into a technique known asDiffusing Wave Spectroscopy (DWS). An important concern for thistechnique is that the analysis is inherently complicated and makesinterpretation of data highly challenging. The agreement of dataobtained from DWS with mechanical data is in many cases quite poor andrequires resealing.

SUMMARY OF THE INVENTION

Several aspects of the invention are disclosed in this application.

Instruments according to the invention can be advantageous in that theycan allow for advanced rheological characterization on very small samplevolumes. They can also allow access to very high-frequency (short time)dynamics.

Instruments according to the invention may allow for improvements inviscoelasticity measurements in a variety of application areas These caninclude high-frequency rheological characterization of complex fluidsfor academic research, personal care, chemicals, and foods, whereinstruments according to the invention can provide an alternative topiezoelectric approaches (PAV/PRV) and DWS. Instruments according to theinvention can also be used in life sciences applications, such as foradvanced rheological characterization of proteins and other biopolymersin solution. In the field of chemicals and specialty chemicals,instruments according to the invention may be used for advancedrheological characterization of newly synthesized polymers or otherchemicals.

Instruments according to the invention may also be used inhigh-throughput applications in a variety of areas, such as academic andpharmaceutical research, personal care, and chemicals.

BRIEF DESCRIPTION OF THE DRAWING

FIG. 1 is a block diagram of a microrheological fluid characteristicmeasurement instrument according to the invention;

FIG. 2 is a plot of an illustrative correlation function for theinstrument of FIG. 1;

FIG. 3 is a plot of DLS-based microrheology data for a 0.5% by weightPEO solution using the instrument of FIG. 1, with mechanical rheometryresults also shown for the same sample; and

FIG. 4 is a plot of DLS-based microrheology data for a 1.0% by weightPEO solution using the instrument of FIG. 1, with mechanical rheometryresults also shown for the same sample.

DETAILED DESCRIPTION OF AN ILLUSTRATIVE EMBODIMENT

Referring to FIG. 1, an illustrative embodiment of a microrheologicalfluid characteristic measurement instrument 10 according to theinvention includes a coherent light source 12, such as a laser, a samplecell 14, at least one detector 20, and a correlator 22. The correlatorcan include autocorrelation logic embodied in hardware and/or softwareto apply an autocorrelation function to a signal from the detector.Single-scattering analysis, such as the viscoelasticity parameterderivations described above, can then be applied to results of thecorrelation operation to extract one or more fluid parametercharacteristics for the sample. An illustrative correlation function isshown in FIG. 2.

The sample cell 14 can be a short-path-length cell, such as a capillarytube having a diameter of 1.5 mm or less. The use of suchshort-path-length cells allows the instrument to minimize multiplescattering contributions to the correlation function in the transmissiongeometry. It is also beneficial in that it allows the instrument to makemeasurements based on small sample amounts, which is particularlyimportant for biomolecules, such as proteins and small-molecule drugs,for which samples can be particularly small. This can allow theinstrument to be used as part of a high-throughput screening system.

The instrument can perform forward-scattering measurements, backscattermeasurements, or both. The use of backscatter detection usingNon-Invasive Back-Scatter (NIBS) techniques can also help to minimizeeffect of multiple scattering contributions to the correlation function.This technique involves performing backscattering measurements at closeto 180°, (e.g., 173°), and is described in U.S. Pat. No. 6,016,195,German patent 19725211, and Japanese patent no. 2911877, which areherein incorporated by reference. The exact NIBS detector spacing andangles will depend on a variety of factors, including the nature of thesample, the material used for the sample vessel, and the desiredaccuracy.

The instrument 10 can also include a fibre 16, a splitter 18, such as a50:50 splitter, and a second detector 20A. The correlator 24 can includecross-correlation logic that allows the instrument to perform across-correlation between the signals from the two detectors. Thiscorrelation operation allows the instrument to more accurately extract aparticle size for samples which are poor scatterers and or are small (afew nm) in size because the effect of the detector dead time, whichdetermines the shortest autocorrelation time, will be reduced. Thecross-correlation operation is also beneficial because it is lesssensitive to detector noise issues, such as afterpulsing, which areuncorrelated between the detectors. And it can allow the correlator todirectly determine the zero time correlation (intercept) of thecorrelation function, improving the calculation of the high frequency G′and G″.

As discussed above, instruments according to the invention can be usedas part of different kinds of high-throughput screening systems. Suchsystems generally include large-scale sample management systems, such asones that are based on scanning mirrors or robotic X-Y stages. TheMalvern Zetasizer APS, for example, provides off-the-shelf automatedmeasurements of samples in industry standard 96- or 384-well plates. Todetect bulk properties of the fluids, the sample vessels should have acapacity that is substantially larger than a domain size of the complexsample fluid and is sufficiently large to cause bulk scattering effectsto substantially exceed surface effects for the complex fluid sample.Exact sample vessel volumes depend on a variety of factors, includingthe nature of the sample and desired accuracy levels.

Instruments according to the invention can be configured to allowscattered light to be detected over a range of different angles, such asfrom 173° to 13.5°. They can also be configured to allow measurements tobe carried out in both backscattering mode or transmission mode in orderto obtain an extended region of frequency response. These objectives canbe accomplished in different ways, such as by allowing a single detectorto move or by providing more than one detector. Measurements can also becarried out using a range of different probe sizes ranging from 30 nm to1 urn in order to extend obtained frequency and/or minimise multiplescattering by adjusting a volume of required probe particles. Andmeasurements can be carried out using a range of different probechemistries to minimise interactions with a complex fluid of interest.

Example 1

In order to validate the above approach, DLS-based optical microrheologywas carried out on the Zetasizer Nano (Malvern Instruments Limited)without any hardware modifications. It should be noted that theZetasizer Nano is designed to implement the NIBS based technique. TheZetasizer Nano is described, for example in U.S. provisional applicationNo. 61/206,688, which is herein incorporated by reference.

The system investigated was a 2M molecular weight PEO (PolyethyleneOxide) formulation at a number of different concentrations. This systemwas already quite turbid even at low concentrations (0.5 wt %) and wascontributing significantly to the scattered light signal. In order toensure domination of the scattering by the probe particles (700 nmnominal diameter, Polystyrene particles, Duke Scientific) they wereadded in relatively high concentrations, which took the system into themoderately multiply scattered regime. As the samples were visibly quiteturbid, measurements would likely have been very difficult to carry outfor them using traditional DLS-based microrheology.

FIGS. 3 and 4 illustrate microrheological data obtained from theZetasizer and comparable data for a high end rotational mechanicalrheometer (Bohlin Gemini HR Nano, Malvern Instruments Limited). The dataclearly illustrates very good agreement with the mechanical data at thelow frequency overlap region and also illustrates the extensivefrequency range over which the viscoelastic response was obtained. TheDLS-based high-frequency data adequately captures the physics (Zimm andRouse Dynamics) that is expected for this system.

In this example, the correlation operation is performed on board theinstrument in a dedicated DSP board and single-scattering analyses areperformed using specialized software running on a general-purposeworkstation. The instrument can also use other approaches to performthese operations, such as dedicated hardware or a combination ofsoftware and dedicated hardware.

The present invention has now been described in connection with a numberof specific embodiments thereof. However, numerous modifications whichare contemplated as falling within the scope of the present inventionshould now be apparent to those skilled in the art. Therefore, it isintended that the scope of the present invention be limited only by thescope of the claims appended hereto. In addition, the order ofpresentation of the claims should not be construed to limit the scope ofany particular term in the claims.

1-15. (canceled)
 16. A fluid characterization measuring instrument,comprising: a sample vessel for a sample fluid, a coherent light sourcepositioned to illuminate the sample fluid in the sample vessel, a firstphoton-counting detector positioned to receive a first portion ofscattered light received from the sample, a second photon-countingdetector positioned to receive a second portion of scattered lightreceived from the sample, cross-correlation logic responsive to thefirst photon-counting detector and to the second photon-countingdetector, and single-scattering fluid property analysis logic responsiveto the cross-correlation logic and operative to derive at least onefluid property for the sample fluid.
 17. The instrument of claim 16wherein the instrument is constructed and adapted to allow the firstand/or second photon-counting detector to allow scattered light to bedetected over a range of different angles ranging from 173° to 13.5°.18. The instrument of claim 16 wherein the instrument is constructed andadapted to allow the first and/or second photon-counting detector to befurther responsive to forward scattered light.
 19. The instrument ofclaim 16 further comprising a beam splitter that splits the coherentlight source into first and second light source portions.
 20. Theinstrument of claim 16 wherein the first photon-counting detector ispositioned to receive the scattered light through a first optical fiberand the second photon-counting detector is positioned to receive thescattered light through a second optical fiber.
 21. The instrument ofclaim 16 wherein the sample vessel is a capillary tube.
 22. Aviscoelasticity measuring method, including the steps of: illuminating asample fluid with coherent light, detecting photons from a first portionof scattered light received from the sample, detecting photons from asecond portion of scattered light received from the sample, performing across-correlation operation between a first detection signalrepresentative of the backscattered photons in the first portion and asecond detection signal representative of the backscattered photons inthe second portion, and deriving at least one fluid property fromresults of the cross-correlation operation for the sample fluid based onsingle-scattering analysis.
 23. The method of claim 22 wherein thecross-correlation operation is an autocorrelation operation.
 24. Themethod of claim 22 wherein scattered light is detected over a range ofdifferent angles ranging from 173° to 13.5° and wherein the steps ofperforming a cross-correlation operation and deriving are performed forthe light detected over a range of angles.
 25. The method of claim 22wherein steps of detecting are carried out in both backscattering modeand forward transmission mode in order to obtain an extended region offrequency response.
 26. The method of claim 22 wherein steps ofdetecting are carried out using a range of different probe sizes rangingfrom 30 nm to 1 μm in order to extend obtained frequency and/or minimisemultiple scattering by adjusting volume of required probe particles. 27.The method of claim 22 wherein steps of detecting are carried out usinga range of different probe chemistries to minimise interactions with thecomplex fluid of interest.
 28. The method of claim 22 whereinilluminating the sample fluid with coherent light comprises splitting acoherent light source into first and second light source portions. 29.The method of claim 22 wherein illuminating the sample fluid withcoherent light comprises illuminating the sample fluid with coherentlight from first and second light sources.
 30. A fluid characterizationmeasuring instrument, comprising: a sample vessel for a sample fluid, afirst coherent light source positioned to illuminate the sample fluid inthe sample vessel, a second coherent light source positioned toilluminate the sample fluid in the sample vessel, a firstphoton-counting detector positioned to receive a first portion ofscattered light received from the sample, a second photon-countingdetector positioned to receive a second portion of scattered lightreceived from the sample, cross-correlation logic responsive to thefirst photon-counting detector and to the second photon-countingdetector, and single-scattering fluid property analysis logic responsiveto the cross-correlation logic and operative to derive at least onefluid property for the sample fluid.
 31. The instrument of claim 30wherein the instrument is constructed and adapted to allow the firstand/or second photon-counting detector to allow scattered light to bedetected over a range of different angles ranging from 173° to 13.5°.32. The instrument of claim 30 wherein the instrument is constructed andadapted to allow the first and/or second photon-counting detector to befurther responsive to forward scattered light.
 33. The instrument ofclaim 30 wherein the first photon-counting detector is positioned toreceive the scattered light through a first optical fiber and the secondphoton-counting detector is positioned to receive the scattered lightthrough a second optical fiber.